流式细胞术做细胞周期分析 | Cell Cycle Analysis by Flow Cytometry
属于几大functional assays中的一个:
- Negative-selection competition assay
- CTG - proliferation
- colony formation assay
- cell cycle
- xenograft
如果你要深入研究一个oncogene或者TSG,你就必须确定其对cancer or tumor viability的影响,如果连表型都没有影响,后面的分子也就没有必要做下去了。
Figure参考:Figure 2.B
We evaluated the role of KAT6A in MLL-r AML in more depth. Consistent with the proliferation phenotypes, we found that loss of KAT6A induced cell-cycle arrest (Fig. 2B) and moderately elevated apoptosis levels in the MLL-r cell lines MOLM-13 and MV4;11 (Supplementary Fig. S2B).
Cell cycle arrest is a stopping point in the cell cycle, where it is no longer involved in the processes surrounding duplication and division.
细胞周期阻滞是指细胞周期停留在G1期,而无法有效的完成整个细胞周期。
G1-phase arrest of cell cycle progression provides an opportunity for cells to either undergo repair mechanisms or follow the apoptotic pathway. The tumor suppressor TP53 plays an important role in response to DNA damage and other genomic instability.
常见的检测方法:
Cell Cycle Analysis by Flow Cytometry
Flow cytometric analysis of cell cycle with propidium iodide DNA staining
需要用到的试剂:
- Propidium Iodide (PI)/RNase Staining Solution
- (Cell Signaling Technology (valid) # 4087S )
为什么不用EdU cell cycle?因为这和CTG的表型是一样的,重叠了。Prytusha的说法。
Click-iT EdU Flow Cytometry Cell Proliferation Assay - 图更fancy一些
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